DNA profiling of disputed chilli samples (Capsicum annum) using ISSR-PCR and FISSR-PCR marker assays

A case of marketing of spurious seeds of chilli. Capsicum annum in the bran d name of an elite variety referred to us from an Indian court of law, for identification is described here. The highly reproducible molecular marker assays, inter simple sequence repeat polymerase chain reaction [ISSR-PCR] a nd FISSR-PCR (for fluorescent ISSR-PCR) were used for differentiating the f our disputed chilli samples. A total number of 17 ISSR anchored primers, wh ich included nine di-, and eight tri-nucleotide primers were used for the a nalysis. The ISSR-PCR products were separated on a 2% agarose gel. A total of 212 and 288 bands were resolved by seven di- and eight tri-nucleotide pr imers, respectively, with an average of 30 bands per primer. Five out of ni ne dinucleotide primers and four out of eight trinucleotide primers could u nambiguously differentiate all the four disputed chilli samples. The sensit ivity and informativeness of the ISSR-PCR assay were further enhanced by th e use of FISSR-PCR technique. The FISSR-PCR assay revealed a total number o f 566 bands using three tri- and one di-nucleotide primers with an average of 141 bands per primer. These four primers could reliably distinguish all the four disputed samples unambiguously. In developing countries like India , violation of Plant Breeder's Rights is a major concern of law. The presen t report is, therefore, a step to protect the Plant Breeder's Rights by mak ing use of reliable and modern DNA technologies. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.